This paper reports the asymmetric separation of atenolol (AT) enantiomers using n-hexyl (L)-tartrate (LT) and boric acid (BA) as chiral selector in a cascade of centrifugal contactor separators (CCSs). A multistage extraction model governing phase equilibrium equations and mass balance equations was established. The model was applied to predict and optimize the asymmetric separation of AT enantiomers. The effects of important factors on the multistage separation process, including the extract to wash phase ratio (0/W), the feed to wash phase ratio (F/W), the concentrations of LT and BA, and the location of feed stage, were simulated and optimized by the multistage extraction model. Under the optimal conditions for asymmetric separation, ATs was purified with enantiomeric excess (ee) of 0.9416 when F/W was 0.50.